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Description

ISOLATE II Plant DNA Kit provides a simple, efficient column-based method for the isolation of genomic DNA from a wide variety of plant materials, without the need for hazardous reagents such as phenol

Product Highlights

Fast – streamlined protocol for reliable recovery of genomic DNA in as little as 30 minutes

High-performance – extraction of high-quality DNA, ideal for use in all downstream applications

Convenient – kit includes all necessary components, including filters (shredders) and RNase A

Safe - no hazardous phenol/chloroform extraction

Product Description

By combining the stringency of CTAB lysis with the speed and ease-of-use of silica-membrane purification, ISOLATE II Plant DNA Kit provides a fast method for the purification of high-quality genomic DNA from most plant cells, particularly those rich in polysaccharides, including leaves, bark, roots and fruits as well as dung, animal-fecal, soil and compost samples.

Some plant tissues do not lyse well in CTAB and so an SDS-based lysis buffer is also provided as an alternative. Residual amounts of RNA remaining can be removed using RNase treatment during lysis with the RNase A supplied in the kit.

ISOLATE II Plant DNA Kit shows excellent recovery of plant DNA when different homogenization techniques are used. The silica membrane in the columns is optimized to improve DNA binding to give high-yields of high-quality DNA, even from difficult samples such as freeze-dried budding leaves. The extracted DNA is ready to use for subsequent downstream reactions such as PCR.

Isolating DNA from soil samples is a big challenge because samples are full of inhibitors like polymers or low-molecular substances, which inhibit downstream applications. Therefore, it is difficult to find a kit which can deliver stable DNA. The ISOLATE II Plant DNA Kit is able to handle these kinds of samples and worked well for soil samples.

Shipping conditions

FAQs

The columns supplied with the ISOLATE II kits may appear similar, but each type has been optimized to work within the buffer system supplied with the corresponding kit. The swapping of columns (or buffers) between kits may lead to no recovery of nucleic acid whatsoever, or at the very least severely impaired purification.

Interruption of the DNA/RNA extraction process is possible after the sample lysis only. It is possible to homogenize and lyse the samples and to store them in the freezer until use for RNA extraction. RNA clean up with a column cannot be interrupted and we recommend to avoid delays during the column purification process. If a delay is unavoidable the columns should be stored on ice.

Both lysis buffers PA1 and PA2 can possibly become cloudy, if they are stored under 20°C. They contain CTAB (PA1) and SDS (PA2) which can precipitate. After a short incubation at 37°C for several minutes they should become clear again.

The ISOLATE II Genomic DNA Kit provides a protocol for hard to lyse bacteria (see sec 9.2). Also the ISOLATE II Plant DNA kit could be suitable, because the optimized lysis buffer from this kits may be also be used for processing these difficult to lyse bacteria. In general, it is necessary to optimize the lysis step with the appropriate mechanical, enzymatic or chemical methods recommended for this kind of sample. Please refer to the related literature or ask the Bioline technical support for further suggestion.

For the optimal recovery of large RNA/DNA fragments it is necessary to optimize the elution step. To increase the recovery rate it would be helpful to use a larger volume for elution, incubate the column with the elution buffer prior to centrifugation and use repeated elution steps. To avoid excessive dilution it may be helpful to reapply the eluted fraction again. Furthermore it could be helpful for DNA elution to heat the elution buffer to 70°C.

Reviews

""Isolating DNA from soil samples is a big challenge because samples are ... "

Katharina Schulte, Technical College Zweibruecken, Germany

""Working with fungi means to accept compromises when using DNA ... "

Karlsruhe, Institute of Technology, Germany

"Working with fungi means to accept compromises when using DNA isolation kits because there aren't specialized kits for fungal application in most cases. The ISOLATE II Plant DNA Kit worked very well for Alternaria alternata and for unknown fungal soil isolates."

Karlsruhe, Institute of Technology, Germany

"Isolating DNA from soil samples is a big challenge because samples are full of inhibitors like polymers or low-molecular substances which inhibit downstream applications. Therefore it is difficult to find a kit which can deliver stable DNA. The ISOLATE II Plant DNA Kit is able to handle these kinds of samples and worked well for soil samples."